The goal of this project is to develop a high-level multiplex real time polymerase chain reaction (PCR) assay for single nucleotide polymorphisms (SNP) using IQuum's innovative Lab-in-a-tube (Liat TM) technology. Liat technology is capable of extracting genomic DNA from whole blood and detecting sequences, using real time PCR, in 30 minutes. Liat technology is ideal for the point of care because it reduces the complexity of nucleic acid tests to a minimum. Indeed, the operator of the Liat Molecular Analyzer, the instrument that performs this genomic DNA sequence detection assay, is only required to input blood into the Liar Tubule and insert the closed Tubule into the instrument. The Liat Molecular Analyzer can also perform multi-color fluorescent probe melting assays in 8 minutes. In Phase I, we propose to investigate the applicability of the molecular inversion probe assay to real time PCR detection using multi-color fluorescent probe melting as a means of detecting multiple SNPs. In addition, we propose to develop a set of hybridization tags with defined melting profiles in order to maximize the multiplexing of this assay. Traditional multiplex real time PCR can only accommodate up to 2 SNPs in one assay. Our strategy will allow us to score up to 40 SNPs in a single assay. We estimate this order of magnitude increase in multiplexing will be accomplished with a 1 hour assay that can be deployed at the point-of-care. In Phase II, we plan to develop a multi-tube Liat Molecular Analyzer that will have the capability of scoring 1.5 million genotypes per year, at the point-of-care. In addition, we propose to develop assays for scoring 40 SNPs that have been associated with increased risk of arterial thrombosis. Venous thrombosis occurs in 0.1 percent of the population each year, and its most severe complication (pulmonary embolism) is as frequent as strokes in developed countries. Arterial thrombosis is a key component of myocardial infarction, stroke and peripheral artery disease, all of which account for >19 million deaths annually. In Phase III, we would collaborate with researchers at Harvard Medical School and Tufts Medical School on clinical trials to evaluate the risk associated with the markers.